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中国癌症防治杂志

中国癌症防治杂志 ›› 2018, Vol. 10 ›› Issue (6): 427-433.doi: 10.3969/j.issn.1674-5671.2018.06.02

• 基础研究 • 上一篇    下一篇

过表达THBS2对宫颈鳞癌SiHa细胞生物学特性的影响

  

  1. 广西医科大学附属肿瘤医院妇瘤科；广西医科大学研究生院
  • 出版日期:2018-12-25 发布日期:2019-01-29
  • 通讯作者: 姚德生 E-mail：yaodesheng@gxmu.edu.cn
  • 基金资助:

    国家自然科学基金资助项目（81460398）；广西自然科学基金资助项目（2017GXNSFAA198106，2015jjAA40619）

Effects of thrombospondin-2 overexpression on the biological behaviors of human cervical squamous carcinoma SiHa cells

  • Online:2018-12-25 Published:2019-01-29

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摘要：

目的 探讨血小板反应蛋白2（thrombospondin-2，THBS2）对人宫颈鳞癌SiHa细胞增殖、侵袭、迁移能力及裸鼠成瘤能力的影响。方法 用THBS2过表达慢病毒转染宫颈鳞癌SiHa细胞（THBS2-LV组）、空载病毒转染宫颈鳞癌SiHa细胞（NC组），以正常培养的宫颈鳞癌SiHa细胞为空白对照组。转染后采用Western blot检测THBS2蛋白的表达，RT-qPCR检测THBS2 mRNA的表达，CCK-8法检测细胞增殖能力，流式细胞术检测细胞凋亡情况，Transwell实验和划痕实验检测细胞侵袭、迁移能力，裸鼠成瘤实验观察各组裸鼠皮下成瘤能力。结果 转染后，与NC组和空白对照组比较，THBS2-LV组细胞THBS2 mRNA和蛋白的表达量升高，细胞增殖、侵袭、迁移能力降低，而细胞凋亡率升高（P＜0.05）。裸鼠体内成瘤实验结果显示，接种THBS2-LV组细胞的裸鼠皮下成瘤能力和平均肿瘤体积均低于接种NC组和空白对照组细胞的裸鼠（P＜0.01）。结论 过表达THBS2能抑制宫颈鳞癌SiHa细胞增殖、侵袭、迁移和裸鼠皮下成瘤能力，THBS2表达上调能抑制宫颈鳞癌发生发展。

关键词: 子宫肿瘤, 宫颈鳞癌, SiHa细胞, 血小板反应蛋白2, 增殖, 凋亡, 体外实验, 动物实验

Abstract:

Objective To investigate the effect of thrombospondin-2（THBS2） on proliferation，invasion and migration of human cervical squamous carcinoma SiHa Cells and the ability of tumor formation in nude mice. Methods Human cervical squamous  carcinoma SiHa cells transfected with the THBS2 overexpression  lentivirus served as the experimental group（THBS2-LV group），transfected with empty virus served as a negative control group （NC group） and untreated human cervical squamous  carcinoma SiHa cells served as a blank control group.The mRNA and THBS2 protein expression were detected by RT-qPCR and Western blot respectively. The proliferation and apoptotic rate of cells were assayed by CCK-8 method and flow cytometry respectively. Cell invasion and migration were analyzed by Transwell assay and Scratch test. The tumor formation ability of nude mice was observed by tumor formation experiment in nude mice. Results Compared with the NC group and the blank control group，the expression of THBS2 mRNA and protein and the apoptotic rate of cells increased in THBS2-LV group，while the cell proliferation，invasion and migration were reduced（P<0.05）. The results of tumor formation in nude mice showed that the subcutaneous tumorigenic ability and average tumor volume of nude mice inoculated with THBS2-LV group were lower than those of nude mice inoculated with NC group and blank control group（P<0.01）. Conclusions Overexpression of THBS2 can inhibit the proliferation，invasion，migration and subcutaneous tumor formation of cervical squamous carcinoma SiHa cells. Up-regulation of THBS2 can inhibit the development of cervical squamous cell carcinoma.

Key words: Uterus neoplasms, Cervical squamous cell carcinoma, SiHa cells, Thrombospondin-2, Proliferation, Apoptosis, Vitro experi-ment, Animal experiment